Dissection of affinity captured LINE-1 macromolecular complexes

Martin S. Taylor, Ilya Altukhov, Kelly R. Molloy, Paolo Mita, Hua Jiang, Emily M. Adney, Aleksandra Wudzinska, Sana Badri, Dmitry Ischenko, George Eng, Kathleen H. Burns, David Fenyö, Brian T. Chait, Dmitry Alexeev, Michael P. Rout, Jef D. Boeke, John LaCava

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)

Abstract

Long Interspersed Nuclear Element-1 (LINE-1, L1) is a mobile genetic element active in human genomes. L1-encoded ORF1 and ORF2 proteins bind L1 RNAs, forming ribonucleoproteins (RNPs). These RNPs interact with diverse host proteins, some repressive and others required for the L1 lifecycle. Using differential affinity purifications, quantitative mass spectrometry, and next generation RNA sequencing, we have characterized the proteins and nucleic acids associated with distinctive, enzymatically active L1 macromolecular complexes. Among them, we describe a cytoplasmic intermediate that we hypothesize to be the canonical ORF1p/ORF2p/L1-RNAcontaining RNP, and we describe a nuclear population containing ORF2p, but lacking ORF1p, which likely contains host factors participating in target-primed reverse transcription.

Original languageEnglish
Article number30094
Number of pages40
JournaleLife
Volume7
DOIs
Publication statusPublished - 8 Jan 2018

Keywords

  • Chromatography, Affinity
  • Endonucleases/analysis
  • HeLa Cells
  • Humans
  • Long Interspersed Nucleotide Elements
  • Macromolecular Substances/chemistry
  • Mass Spectrometry
  • RNA-Directed DNA Polymerase/analysis
  • RNA/analysis
  • Ribonucleoproteins/analysis
  • HOST FACTORS
  • REVERSE TRANSCRIPTION
  • L1 RETROTRANSPOSITION
  • BINDING-PROTEINS
  • MESSENGER-RNA
  • REPLICATION FORKS
  • AMINO-ACIDS
  • ORF1 PROTEIN
  • CELL-CULTURE
  • STRESS GRANULES

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